Biological properties of necrotic and non-necrotic strains of bean yellow mosaic virus in cool season grain legumes

A collection of 51 bean yellow mosaic virus (BYMV) isolates was transmitted from infected Trifolium subterraneum (subterranean clover) to Lupinus angustifolius (narrow‐leafed lupin) by Myzus persicae (green peach aphid). Depending on isolate and L. angiistifolius genotype used, two distinct responses developed in L. angustifolius plants, either systemic necrosis and plant death or non‐necrotic reactions of varying severity. Ten isolates caused necrosis and plant death in cv. Danja. However, when nine of these were inoculated to breeding line 90L423‐07‐13, seven induced non‐necrotic reactions, while two caused necrosis and plant death. Thirty seven isolates always produced non‐necrotic reactions regardless of genotype of L. angustifolius inoculated. Non‐necrotic and necrotic isolates originally came both from lupins and other species, and the non‐necrotic isolates were no less efficiently transmitted by M. persicae than the necrotic ones. When one isolate of each type was inoculated together to T. subterraneum and nine months later this culture was used as an acquisition source for aphid transmission to L. angustifolius, only the necrotic type was detected. Previous infection of L. angustifolius plants with a non necrolic isolate prevented subsequent infection by a necrotic one. All necrotic and non‐necrotic isolates reacted with BYMV antiserum in ELISA but only two cross‐reacted with antiserum to clover yellow vein virus (CYVV). When selected necrotic and non‐necrotic isolates were inoculated to differential hosts, all behaved like BYMV and not CYVV. When three isolates of each type were transmitted to 11 other cool season grain legume species, except in Cicer arietinum (chickpea), there were no necrotic reactions, but symptom severity varied with the isolate and species inoculated. The two isolates that caused necrosis in C. arietinum did not do so in L. angustifolius. The six isolates from Vicia faba (faba bean) all caused non‐necrotic reactions in L. angustifolius cv. Danja and 90L423‐07‐13. These and two necrotic isolates readily infected five genotypes of V. faba always causing severe symptoms. However, three non‐necrotic isolates from L. angustifolius and a further necrotic isolate were poorly infectious on V. faba in which they generally induced mild symptoms. These results show that at least three strain groups of BYMV can be distinguished by their reactions in different L. angustifolius genotypes, one causing necrosis and death in cv. Danja and 90L423‐07‐13, one causing necrosis and death in Danja but not 90L423‐07‐13, and one causing non‐nccrotic reactions in both. These strain groups could not be distinguished when representative necrotic and non‐necrotic isolates were inoculated to other grain legume species. However, inoculation to V. faba distinguished two other BYMV strain groupings differing in severity of symptoms and ability to infect this species.     

Selection of resistance breaking strains of tomato spotted wilt tospovirus

In glasshouse tests, sap from plants infected with 15 different isolates of tomato spotted wilt tospovirus (TSWV) from three Australian states was inoculated to nine genotypes of tomato carrying TSWV resistance gene Sw-5 or one of its alleles. A further two resistant tomato genotypes were inoculated with four isolates each. The normal response in resistant genotypes was development of necrotic local lesions in inoculated leaves without systemic invasion, but 22/752 plants also developed systemic reactions in addition to local hypersensitive ones. Using cultures from two of these systemically infected plants and following four cycles of subculture in TSWV resistant tomato plants, two isolates were obtained that gave susceptible type systemic reactions but no necrotic spots in inoculated leaves of resistant tomatoes. When these two isolates, Da_WA-1d and To_TAS-1d, were maintained by repeated subculture for 10 successive cycles in Nicotiana glutinosa or a susceptible tomato genotype, they still induced susceptible type systemic reactions when inoculated to resistant tomato plants. They were therefore stable resistance breaking isolates as regards overcoming gene Sw-5. When resistance-breaking isolate Da_WA^-1ld multiplied together with original isolate Da_WA-l in susceptible tomato, it was fully competitive with the original isolate. However, when Da_WA-ld and To_TAS-ld were inoculated to TSWV resistant Lycopersicon peruvianum lines PI 128660R and PI 128660S and to TSWV resistant Capsicum chinense lines PI 152225, PI 159236 and AVRDC CO0943, they failed to overcome the resistance, producing only necrotic local lesions without systemic infection. Thus, although the ease of selection, stability and competitive ability of resistance breaking isolates of TSWV is cause for concern, L. peruvianum and C. chinense lines are available which are effective against them. The effectiveness of the resistance to TSWV in nine tomato genotypes was examined in a field experiment. Spread was substantial in the susceptible control genotype infecting 42% of plants. Resistance was ineffective in cv. Bronze Rebel, 26% of plants developing infection. In contrast, it held up well in the other eight resistant genotypes with only 1–3 or no plants of each becoming infected. Accumulated numbers of Thrips tabaci, Frankliniella occidentalis and F. schultzei were closely correlated with TSWV spread.     

The effect of different virus isolates on the expression of tolerance to barley yellow dwarf virus in barley

A barley variety of Ethiopian origin, with a single Mendelian gene con-fering tolerance to barley yellow dwarf virus (BYDV), was equally tolerant to a number of isolates of the virus, whereas a susceptible European barley was more susceptible to isolates transmitted by Rhopalosiphum padi L. than to those transmitted by Macrosiphum (Sitobion) avenae (Fab). However, hybrids between these two varieties homozygous for the Ethiopian tolerance gene were more tolerant to ‘mild’ than to ‘severe’ isolates, irrespective of the vector specificity. The European variety was damaged more severely by all isolates when infected early than when infected late in its development, but the hybrids were damaged more severely by M. awraae-transmitted isolates when infected late. It is suggested that in susceptible plants the concentration, rather than the virulence, of the virus determines disease severity, whereas the reverse is true in plants possessing a gene which reduces virus multiplication. Virus concentration appears to determine the severity of R. padi-transmitted isolates, while virulence determines the severity of M. avenae-transmitted isolates. The latter would also seem to be adapted towards late infection.     

Mechanism of paraquat tolerance in perennial ryegrass

Abstract The mechanism of paraquat tolerance was investigated in lines of perennial ryegrass (Lolium perenne L.) which had been selected for resistance to the herbicide. Uptake, metabolism and translocation of paraquat were studied. Susceptible cultivars and a tolerant line were not found to differ in uptake of radioactive paraquat applied to the leaf surface or supplied to the cut ends of excised leaves. Distribution of herbicide within leaf tissue was similar in tolerant and susceptible plants and no metabolites of ¹⁴C-paraquat were detected in tolerant or susceptible plants treated with sub-lethal concentrations of the herbicide. Autoradiography and quantitative determinations showed much variation in translocation of ¹⁴C-paraquat out of treated leaves of intact plants, but the variation was not related to the degree of susceptibility to the herbicide. It is concluded that paraquat tolerance in perennial ryegrass is unlikely to depend upon reduced uptake, enhanced metabolism or altered translocation of the herbicide.     

A necrosis-inducing elicitor domain encoded by both symptomatic and asymptomatic Plantago asiatica mosaic virus isolates, whose expression is modulated by virus replication

Systemic necrosis is the most destructive symptom induced by plant pathogens. We previously identified amino acid 1154, in the polymerase domain (POL) of RNA-dependent RNA polymerase (RdRp) of Plantago asiatica mosaic virus (PlAMV), which affects PlAMV-induced systemic necrosis in Nicotiana benthamiana. By point-mutation analysis, we show that amino acid 1,154 alone is not sufficient for induction of necrotic symptoms. However, PlAMV replicons that can express only RdRp, derived from a necrosis-inducing PlAMV isolate, retain their ability to induce necrosis, and transient expression of PlAMV-encoded proteins indicated that the necrosis-eliciting activity resides in RdRp. Moreover, inducible-overexpression analysis demonstrated that the necrosis was induced in an RdRp dose-dependent manner. In addition, during PlAMV infection, necrotic symptoms are associated with high levels of RdRp accumulation. Surprisingly, necrosis-eliciting activity resides in the helicase domain (HEL), not in the amino acid 1,154-containing POL, of RdRp, and this activity was observed even in HELs of PlAMV isolates of which infection does not cause necrosis. Moreover, HEL-induced necrosis had characteristics similar to those induced by PlAMV infection. Overall, our data suggest that necrotic symptoms induced by PlAMV infection depend on the accumulation of a non-isolate specific elicitor HEL (even from nonnecrosis isolates), whose expression is indirectly regulated by amino acid 1,154 that controls replication.     

Tolerance to ryegrass mosaic virus, its assessment and effect on yield

Symptom severity of eighteen populations of Italian ryegrass infected artificially in the glasshouse and naturally in the field with ryegrass mosaic virus (RMV) was strongly correlated. A smaller proportion of plants of the more tolerant populations showed symptoms in the field, but this was probably due to an association of tolerance with increasing incidence of symptomless infection rather than with resistance to infection. Under sward conditions, the yield of a sensitive genotype was reduced by 27% and that of two more tolerant ones by 15 and 13 %. The percentage dry matter yield loss of the most sensitive genotype was similar in all cuts, despite the appearance of extensive necrosis at the time of one cut. With the more tolerant genotype the yield loss varied from 7 to 25 % according to cut. Over the period of the experiment RMV infection did not increase plant mortality.     

Genotype-specific differences in chilling tolerance of maize in relation to chilling-induced changes in water status and abscisic acid accumulation

Four inbred maize lines differing in chilling tolerance were used to study changes in water status and abscisic acid (ABA) levels before, during and after a chilling period. Seedlings were raised in fertilized soil at 24/22°C (day/night), 70% relative humidity. and a 12-h photoperiod with 200 μmol m⁻² s⁻¹ from fluorescent tubes. At an age of 2 weeks the plants were conditioned at 14/12°C for 4 days and then chilled for 5 days at 5/3°C. The other conditions (relative humidity, quantum flux, photoperiod) were unchanged. After the chilling period the plants were transferred to the original conditions for recovery. The third leaves were used to study changes in leaf necrosis, ion efflux, transpiration, water status and ABA accumulation. Pronounced differences in chilling tolerance between the 4 lines as estimated by necrotic leaf areas, ion efflux and whole plant survival were observed. Conditioning significantly increased tolerance against chilling at 5/3°C in all genotypes. The genotypes with low chilling tolerance had lower water and osmotic potentials than the more tolerant genotypes during a chilling period at 5/3°C. These differences were related to higher transpiration rates and lower diffusive resistance values of the more susceptible lines. During chilling stress at 5/3°C ABA levels were quadrupled. Only a small rise was measurable during conditioning at 14/12°C. However, conditioning enhanced the rise of ABA during subsequent chilling. ABA accumulation in the two lines with a higher chilling tolerance was triggered at a higher leaf water potential and reached higher levels than in the less tolerant lines. We conclude that chilling tolerance in maize is related to the ability for fast and pronounced formation of ABA as a protective agent against chilling injury.     

Natural resistance to cucumber mosaic virus in lupin species

Ten species of lupins (Lupinus spp.) were tested for resistance to cucumber mosaic cucumovirus (CMV) in field experiments where inoculation was by naturally-occurring aphid vectors, and in the glasshouse by sap or graft-inoculation. L. albus and six species of ‘rough-seeded’ lupins did not become infected with CMV either under intense inoculum pressure in the field or when graft-inoculated. Two L. hispanicus, 17 L. luteus and four L. mutabilis genotypes became infected with CMV in the field, but no infection was detected in L. hispanicus P26858 or seven L. luteus genotypes. CMV was detected at seed transmission rates of 0.2–16% in seedlings of infected L. luteus, differences in levels of seed transmission between genotypes being significant and relatively stable from year to year. Graft-inoculation of CMV to plants of six genotypes of L. luteus in which no infection was found in the field induced a systemic necrotic reaction suggesting that the resistance they carry is due to hypersensitivity. In L. hispanicus accessions P26849, P26853 and P26858, CMV sub-group II isolate SN caused necrotic spots in inoculated leaves without systemic movement, while sub-group I isolate SL infected them systemically without necrosis. Another sub-group I and two other sub-group II isolates behaved like SL in P26849 and P26853 but infected only inoculated leaves of P26858. This suggests that two strain specific hypersensitive resistance specificities are operating against CMV in L. hispanicus. When plants of L. luteus genotypes that gave hypersensitive reactions on graft-inoculation were inoculated with infective sap containing two sub-group I and seven sub-group II isolates, they all responded like L. hispanicus P26858. A strain group concept is proposed for CMV in lupins based on the two hypersensitive specificities found: strain group 1 represented by isolate SN which induces hypersensitivity with both specificities, strain group 2 represented by the three isolates which induced hypersensitivity only with the specificity present in L. luteus and L. hispanicus P26858, strain group 3 by as yet hypothetical isolates that induce hypersensitivity only in presence of the specificity in L. hispanicus P26849 and P26853 that responded just to isolate SN, and strain group 4 by isolate SL which overcomes both specificities. When F₂ progeny plants from crosses between hypersensitive and susceptible L. luteus parents were inoculated with isolate SN, the resistance segregated with a 3:1 ratio (hypersensitive:susceptible), suggesting that a single dominant hypersensitivity gene, Ncm-1, is responsible. As gene Ncm-1 had broad specificity and was not overcome by any of the five CMV isolates from lupins tested, it is valuable for use in breeding CMV resistant L. luteus cultivars.     

Natural resistance to cucumber mosaic virus in lupin species

Ten species of lupins (Lupinus spp.) were tested for resistance to cucumber mosaic cucumovirus (CMV) in field experiments where inoculation was by naturally-occurring aphid vectors, and in the glasshouse by sap or graft-inoculation. L. albus and six species of ‘rough-seeded’ lupins did not become infected with CMV either under intense inoculum pressure in the field or when graft-inoculated. Two L. hispanicus, 17 L. luteus and four L. mutabilis genotypes became infected with CMV in the field, but no infection was detected in L. hispanicus P26858 or seven L. luteus genotypes. CMV was detected at seed transmission rates of 0.2–16% in seedlings of infected L. luteus, differences in levels of seed transmission between genotypes being significant and relatively stable from year to year. Graft-inoculation of CMV to plants of six genotypes of L. luteus in which no infection was found in the field induced a systemic necrotic reaction suggesting that the resistance they carry is due to hypersensitivity. In L. hispanicus accessions P26849, P26853 and P26858, CMV sub-group II isolate SN caused necrotic spots in inoculated leaves without systemic movement, while sub-group I isolate SL infected them systemically without necrosis. Another sub-group I and two other sub-group II isolates behaved like SL in P26849 and P26853 but infected only inoculated leaves of P26858. This suggests that two strain specific hypersensitive resistance specificities are operating against CMV in L. hispanicus. When plants of L. luteus genotypes that gave hypersensitive reactions on graft-inoculation were inoculated with infective sap containing two sub-group I and seven sub-group II isolates, they all responded like L. hispanicus P26858. A strain group concept is proposed for CMV in lupins based on the two hypersensitive specificities found: strain group 1 represented by isolate SN which induces hypersensitivity with both specificities, strain group 2 represented by the three isolates which induced hypersensitivity only with the specificity present in L. luteus and L. hispanicus P26858, strain group 3 by as yet hypothetical isolates that induce hypersensitivity only in presence of the specificity in L. hispanicus P26849 and P26853 that responded just to isolate SN, and strain group 4 by isolate SL which overcomes both specificities. When F₂ progeny plants from crosses between hypersensitive and susceptible L. luteus parents were inoculated with isolate SN, the resistance segregated with a 3:1 ratio (hypersensitive:susceptible), suggesting that a single dominant hypersensitivity gene, Ncm-1, is responsible. As gene Ncm-1 had broad specificity and was not overcome by any of the five CMV isolates from lupins tested, it is valuable for use in breeding CMV resistant L. luteus cultivars.     

IV. Variation in fungicide tolerance: Comparisons between isolates of Erysiphe graminis f. sp. hordei tolerant and sensitive to the fungicide tridemorph

Forty-five single pustule isolates of barley powdery mildew taken at random from tridemorph treated and untreated plants were maintained in culture on leaf segments of barley cv. Golden Promise placed on agar containing benzimidazole. In repeated assays for tridemorph tolerance over a 10 month period, tolerance was found in 25 of 36 isolates from treated plants, and remained stable in all but four of the isolates wherein tolerance was lost after varying lengths of time. A negative correlation was found between pathogenicity and tolerance. The germination of tolerant isolates was less depressed than the sensitive on treated seedlings, but no differences between isolates were found on untreated seedlings. Colony growth of all isolates was slower on treated than untreated seedlings. On plants of several cultivars kept in growth rooms, mildew progressed more rapidly on treated barley when tolerant isolates were used as inoculum. Visible disease symptoms spread most rapidly on isolated, untreated plants, and most slowly on isolated treated plants, irrespective of the isolate. Disease on treated plants progressed more rapidly when untreated plants were adjacent, suggesting that interactions take place between pathogen populations on treated and untreated plants.     

Resistance to cucumber mosaic virus in potato

Reactions to two subgroup I isolates (Fny-CMV and Pf-CMV) and two subgroup II isolates (A9-CMV and LS-CMV) of cucumber mosaic virus (CMV) were studied in three non tuber-bearing wild potato species (Solanum spp.) of the series Etuberosa, and in two tuber-bearing interspecific potato hybrids and four potato cultivars using graft-inoculation. Three classes of phenotypic reactions (susceptible, hypersensitive, extreme resistance) were observed in the tuber-bearing genotypes. Susceptible genotypes developed mosaic or severe mosaic with leaf malformation and had high CMV titres. Hypersensitive genotypes developed either top necrosis or vein necrosis and/or necrotic spots on apical leaves, and had low CMV titres. Extremely resistant genotypes had no symptoms and no CMV was detected. The hybrid 87HW13.7 (S. tuberosum×S. multidissectum) developed top necrosis specific to infection with Fny-CMV. The hybrid ‘A6’ (S. demissum×S. tuberosum cv. Aquila) was hypersensitive to all CMV isolates tested. Extreme resistance was not functional against all CMV isolates. Neither hypersensitivity nor extreme resistance were related to the CMV subgroup.     

Lettuce mosaic virus pathogenicity determinants in susceptible and tolerant lettuce cultivars map to different regions of the viral genome

Full-length infectious cDNA clones were constructed for two isolates (LMV-0 and LMV-E) of Lettuce mosaic virus (LMV), a member of the genus Potyvirus. These two isolates differ in pathogenicity in susceptible and tolerant-resistant lettuce cultivars. In susceptible plants, LMV-0 induces mild mosaic symptoms, whereas LMV-E induces severe stunting, leaf deformation, and a necrotic mosaic. In plants carrying either of the two probably allelic recessive resistance genes mol1 or mol2, LMV-0 is restricted partially to the inoculated leaves. When a systemic invasion does occur, however, symptoms fail to develop. LMV-E overcomes the protection afforded by the resistance genes, resulting in systemic mosaic symptoms. Analysis of the behavior of recombinants constructed between the two virus isolates determined that the HC-Pro protein of LMV-E causes the severe stunting and necrotic mosaic induced by this isolate in susceptible cultivars. In contrast, the ability to overcome mol resistance and induce symptoms in the resistant-tolerant cultivars was mapped to the 3' half of the LMV-E genome. These results indicate that the ability to induce severe symptoms and to overcome the protection afforded by the recessive genes mol1 or mol2 are independent phenomena.     

Tolerance in Isolates of Colletotrichum coffeanum to Prochloraz-Mn in Kenya

Coffee berry disease, caused by Colletotrichum coffeanum is a serious disease of Coffea arabica in Kenya. Control of this disease is achieved mainly through the use of fungicides which currently include chlorothalonil and prochloraz-Mn (Octave). Fungicide resistance in C. coffeanum to benzimidazoles has been well documented. Isolates of C. coffeanum highly sensitive to prochloraz-Mn have been shown to acquire tolerance to high levels of prochloraz-Mn in culture. These isolates were able to grow and sporulate in malt extract agar (MEA) amended with 250 μg ml^-1 prochloraz-Mn. The prochloraz-Mn tolerant isolates of C. coffeanum showed low level of tolerance to benomyl. The benomyl tolerant isolates of C. coffeanum equally showed low level of tolerance to prochloraz-Mn. The prochloraz-Mn and benomyl tolerant and sensitive isolates were found to be highly pathogenic and induced sporulating lesions on seedlings and berries of coffee cv. SL 28 whichis, very susceptible to C. coffeanum. Mixed inoculation tests using prochloraz-Mn and benomyl tolerant isolates and prochloraz-Mn tolerant and sensitive isolates of C. coffeanum showed that the prochloraz-Mn tolerant isolate was competitive.     

Quantitative differences in aphid virulence and foliar symptom development on tomato plants carrying the Mi resistance gene

The Mi resistance gene in tomato reduces the feeding, fecundity, and survival of certain isolates of the potato aphid (Macrosiphum euphorbiae Thomas). This study compared the performance of two potato aphid isolates, WU11 and WU12, on nearly isogenic susceptible (Mi-) and resistant (Mi+) tomato cultivars. Although Mi significantly reduced the population growth of both aphids, WU12 numbers decreased by only 15% compared with 95% for isolate WU11. These results show that there are quantitative differences in virulence among potato aphid isolates. Compared with WU11 aphids, isolate WU12 caused more necrosis on both resistant and susceptible plants, and this increased damage may play a role in the partial virulence of isolate WU12. However, infestation with aphid isolate WU12 did not compromise plant defenses against isolate WU11 in resistant plants. Prior inoculation with either aphid isolate caused a modest reduction in the survival of WU12 adults, but this form of induced resistance was observed on both resistant and susceptible cultivars. Thus, Mi did not play a role in acquired resistance or mediate any indirect interactions between the two aphid isolates. Notably, the mode of action of Mi-mediated resistance seemed to differ depending on the aphid isolate tested. Mi dramatically deterred feeding by WU11 aphids, whereas the effects of resistance on isolate WU12 seemed to be caused primarily by antibiosis. Tolerance did not seem to be a major component of Mi-mediated responses, although resistant plants showed a modest reduction in the amount of foliar necrosis induced per aphid compared with susceptible plants.     

The effects of rootstock and scion on tobacco mosaic virus infection in susceptible, tolerant and immune cultivars of tomato

Reciprocal grafts were made between tomato cultivars Potentate, susceptible, and Virocross, tolerant (heterozygous for resistance gene Tm-i) to tobacco mosaic virus (TMV) isolates of Pelham type o and between isogenic lines of cv. Craigella, susceptible and homozygous for gene Tm-i. The grafted plants were inoculated with a type o isolate; both scion and stock inoculation were studied in the former, scion inoculation only, in the latter. With scion inoculation the virus content of a tolerant scion was greater on a susceptible stock than on a tolerant one, but that of a susceptible scion was unaffected by the type of stock: in contrast, the virus content of a tolerant stock was unaffected by the type of scion but that of a susceptible stock was less with a tolerant than with a susceptible scion. With root inoculation the virus contents of both tolerant and susceptible scions were greater on a susceptible than on a tolerant stock. With cv. Craigella the genotype Tm-1/Tm-1 was found to be immune to the type o isolate used, but in grafts the leaves of Tm-1/Tm-1 scions became tolerant to leaf inoculation when on susceptible stocks and the virus entered the stock. Tm-1/Tm-1 stocks became infected when attached to infected, susceptible scions and did not affect the virus content of those scions. The results indicate that a susceptible healthy stock may change the reaction of a tolerant or immune scion to infection by a strain of TMV.     

Inheritance of hypersensitive resistance to Bean yellow mosaic virus in narrow-leafed lupin (Lupinus angustifolius)

In narrow‐leafed lupin (Lupinus angustifolius), segregation for the necrotic (systemic hypersensitive) response to infection with a necrotic strain of Bean yellow mosaic virus (BYMV‐N) was studied in progeny plants from six crosses. The parents were two cultivars that always developed necrosis when infected (Danja and Merrit) and two genotypes that always responded without necrosis (90L423‐07‐13 and P26697). In the four possible combinations of crosses between the different necrotic and non‐necrotically reacting genotypes, segregation for the necrotic response in F₂ progeny plants always fitted a 3:1 ratio (necrotic: non‐necrotic). All F₂ progeny plants from the cross between the two non‐cultivar genotypes became infected without necrosis while 99% of the F₂ from the cross between the two cultivars developed necrosis. These results indicate that the systemic necrotic response to infection with BYMV‐N is probably controlled by a single dominant hypersensitivity gene for which we propose the name Nbm‐1. However, its expression seemed influenced by independently segregating modifier genes in the genetic background since necrosis developed at widely different rates within affected F₂ progeny plants resulting in staggered killing.     

The resistance of the perennial ryegrass cultivar Endura to ryegrass mosaic virus

The perennial ryegrass cv. Endura is particularly resistant to infection with two ryegrass mosaic virus isolates, RMV-Roth and RMV-Sax. A few plants appeared immune to RMV-Roth but were infected by RMV-Sax. Two plants that developed only mild symptoms after infection with RMV-Roth contained few virus particles, but passage through these resistant plants caused no detectable change in the virulence of RMV-Roth towards S 22 Italian ryegrass. In offsets derived from one of these resistant plants, RMV-Sax caused severe symptoms and attained a high virus concentration but it was unable to infect if the plant was already infected with RMV-Roth.     

DNA Methylation Alterations and Their Association with High Temperature Tolerance in Rice Anthesis

DNA methylation is an important epigenetic mechanism involved in gene regulation under environmental stresses in plants. However, little information is available regarding its responses to high temperature (HT) and association with HT tolerance in rice. In this study, fourteen rice genotypes were classified into the susceptible, moderate, and tolerant groups by the high temperature susceptibility index (HTSI) after HT treatment. The changes of DNA methylation in rice anthesis under normal and HT30 conditions were investigated using methylation-sensitive amplified polymorphism31 (MSAP). The MSAP results showed that the DNA methylation level significantly increased in the susceptible rice group and decreased in the tolerant rice group under HT treatment, while no significant difference was observed in the moderate rice group. More hypomethylation events were detected in the tolerant rice group, while more hypermethylation was detected in the susceptible rice group. Forty-four differentially methylated epiloci (DME) were generated under both control and HT conditions, which can clearly distinguish the susceptible, moderate, and tolerant genotypes via PCoA analysis. Approximately 43.18% of DMEs were determined to be tolerance-associated epiloci (TAEs). 63.15% TAEs were sequenced and annotated into 12 genes. Quantitative RT-PCR analysis showed that 12 TAE genes were mainly upregulated in 14 rice genotypes, and their expression levels were related to the HT tolerance of rice. Here, DEGs, generated from a number of genotypes, indicate higher probabilities for association with stress tolerance. Overall, these results suggest that DNA methylation regulation might play a key role in adaptation to HT stress in rice.